Document Detail

Identification of a cKit(+) colonic crypt base secretory cell that supports Lgr5(+) stem cells in mice.
MedLine Citation:
PMID:  22333952     Owner:  NLM     Status:  MEDLINE    
BACKGROUND & AIMS: Paneth cells contribute to the small intestinal niche of Lgr5(+) stem cells. Although the colon also contains Lgr5(+) stem cells, it does not contain Paneth cells. We investigated the existence of colonic Paneth-like cells that have a distinct transcriptional signature and support Lgr5(+) stem cells.
METHODS: We used multicolor fluorescence-activated cell sorting to isolate different subregions of colon crypts, based on known markers, from dissociated colonic epithelium of mice. We performed multiplexed single-cell gene expression analysis with quantitative reverse transcriptase polymerase chain reaction followed by hierarchical clustering analysis to characterize distinct cell types. We used immunostaining and fluorescence-activated cell sorting analyses with in vivo administration of a Notch inhibitor and in vitro organoid cultures to characterize different cell types.
RESULTS: Multicolor fluorescence-activated cell sorting could isolate distinct regions of colonic crypts. Four major epithelial subtypes or transcriptional states were revealed by gene expression analysis of selected populations of single cells. One of these, the goblet cells, contained a distinct cKit/CD117(+) crypt base subpopulation that expressed Dll1, Dll4, and epidermal growth factor, similar to Paneth cells, which were also marked by cKit. In the colon, cKit(+) goblet cells were interdigitated with Lgr5(+) stem cells. In vivo, this colonic cKit(+) population was regulated by Notch signaling; administration of a γ-secretase inhibitor to mice increased the number of cKit(+) cells. When isolated from mouse colon, cKit(+) cells promoted formation of organoids from Lgr5(+) stem cells, which expressed Kitl/stem cell factor, the ligand for cKit. When organoids were depleted of cKit(+) cells using a toxin-conjugated antibody, organoid formation decreased.
CONCLUSIONS: cKit marks small intestinal Paneth cells and a subset of colonic goblet cells that are regulated by Notch signaling and support Lgr5(+) stem cells.
Michael E Rothenberg; Ysbrand Nusse; Tomer Kalisky; John J Lee; Piero Dalerba; Ferenc Scheeren; Neethan Lobo; Subhash Kulkarni; Sopheak Sim; Dalong Qian; Philip A Beachy; Pankaj J Pasricha; Stephen R Quake; Michael F Clarke
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2012-02-11
Journal Detail:
Title:  Gastroenterology     Volume:  142     ISSN:  1528-0012     ISO Abbreviation:  Gastroenterology     Publication Date:  2012 May 
Date Detail:
Created Date:  2012-04-24     Completed Date:  2012-06-18     Revised Date:  2014-10-26    
Medline Journal Info:
Nlm Unique ID:  0374630     Medline TA:  Gastroenterology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1195-1205.e6     Citation Subset:  AIM; IM    
Copyright Information:
Copyright © 2012 AGA Institute. Published by Elsevier Inc. All rights reserved.
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MeSH Terms
Antigens, CD / analysis
Antigens, CD44 / analysis
Cell Adhesion Molecules / analysis
Cells, Cultured
Colon / cytology*
Flow Cytometry
Gene Expression Profiling
Goblet Cells / physiology
Mice, Inbred C57BL
Paneth Cells / chemistry*,  physiology*
Proto-Oncogene Proteins c-kit / analysis*
Receptors, G-Protein-Coupled / analysis*
Receptors, Notch / physiology
Single-Cell Analysis
Stem Cells / chemistry,  physiology*
Grant Support
5P01CA139490-03/CA/NCI NIH HHS; 5P30DK056339/DK/NIDDK NIH HHS; P01 CA139490/CA/NCI NIH HHS; R01 CA100225/CA/NCI NIH HHS; R01 CA104987/CA/NCI NIH HHS; R01 DK080920/DK/NIDDK NIH HHS; T32 DK0070560/DK/NIDDK NIH HHS
Reg. No./Substance:
0/Antigens, CD; 0/Antigens, CD44; 0/CD66 antigens; 0/Cell Adhesion Molecules; 0/Lgr5 protein, mouse; 0/Receptors, G-Protein-Coupled; 0/Receptors, Notch; EC Proteins c-kit

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